Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Arm and head domain in highly conserved lipoprotein modification enzyme Lgt determine functional diversity among bacterial pathogens.

mBio·2026
Same author

When the Host Encounters the Cell Wall and Vice Versa.

Annual review of microbiology·2024
Same author

Components Subcellular Localization: Identification of Lipoproteins Using Alkyne Fatty Acids and Click Chemistry.

Methods in molecular biology (Clifton, N.J.)·2023
Same author

Post-translational modifications in bacteria - The dynamics of bacterial physiology.

Research in microbiology·2021
Same author

Mode of action of lipoprotein modification enzymes-Novel antibacterial targets.

Molecular microbiology·2020
Same author

Click-Chemistry Based Fluorometric Assay for Apolipoprotein N-acyltransferase from Enzyme Characterization to High-Throughput Screening.

Journal of visualized experiments : JoVE·2020

Related Experiment Video

Updated: Jul 11, 2025

Quantitative Localization of a Golgi Protein by Imaging Its Center of Fluorescence Mass
13:08

Quantitative Localization of a Golgi Protein by Imaging Its Center of Fluorescence Mass

Published on: August 10, 2017

10.8K

Components Subcellular Localization: Identification of Lipoproteins Using Globomycin and Radioactive Palmitate.

Nienke Buddelmeijer1

  • 1Institut Pasteur, Université Paris Cité, CNRS UMR6047, INSERM U1306, Biology and genetics of the bacterial cell wall Unit, 25-28 rue du docteur Roux, Paris cedex 15, France. nienke.buddelmeijer@pasteur.fr.

Methods in Molecular Biology (Clifton, N.J.)
|November 6, 2023
PubMed
Summary

Researchers detected an intermediate form of bacterial lipoproteins, diacylglyceryl-prolipoprotein. This was achieved by labeling with 3H-palmitate and inhibiting signal peptidase II (Lsp) with globomycin.

Keywords:
3H-palmitate labelingFluorographyGlobomycinTris-Tricine SDS gel electrophoresis

More Related Videos

Using Caco-2 Cells to Study Lipid Transport by the Intestine
07:00

Using Caco-2 Cells to Study Lipid Transport by the Intestine

Published on: August 20, 2015

18.9K
Determination of Plasma Membrane Partitioning for Peripherally-associated Proteins
11:11

Determination of Plasma Membrane Partitioning for Peripherally-associated Proteins

Published on: June 15, 2018

8.4K

Related Experiment Videos

Last Updated: Jul 11, 2025

Quantitative Localization of a Golgi Protein by Imaging Its Center of Fluorescence Mass
13:08

Quantitative Localization of a Golgi Protein by Imaging Its Center of Fluorescence Mass

Published on: August 10, 2017

10.8K
Using Caco-2 Cells to Study Lipid Transport by the Intestine
07:00

Using Caco-2 Cells to Study Lipid Transport by the Intestine

Published on: August 20, 2015

18.9K
Determination of Plasma Membrane Partitioning for Peripherally-associated Proteins
11:11

Determination of Plasma Membrane Partitioning for Peripherally-associated Proteins

Published on: June 15, 2018

8.4K

Area of Science:

  • Microbiology
  • Molecular Biology
  • Biochemistry

Background:

  • Bacterial lipoproteins are essential cell envelope components.
  • Lipoproteins undergo posttranslational modification, including fatty acid acylation.
  • The precise intermediate steps in lipoprotein maturation are not fully understood.

Purpose of the Study:

  • To detect and characterize an intermediate form of bacterial lipoprotein.
  • To investigate the role of signal peptidase II (Lsp) in lipoprotein processing.

Main Methods:

  • Utilized 3H-palmitate labeling to trace fatty acid incorporation.
  • Employed globomycin to specifically inhibit signal peptidase II (Lsp).
  • Applied fluorography for sensitive detection of labeled intermediates.

Main Results:

  • Successfully detected diacylglyceryl-prolipoprotein, an intermediate in lipoprotein synthesis.
  • Demonstrated the utility of Lsp inhibition for isolating precursor forms.
  • Confirmed the covalent attachment of fatty acids to the lipoprotein precursor.

Conclusions:

  • Diacylglyceryl-prolipoprotein is a detectable intermediate in bacterial lipoprotein biogenesis.
  • Signal peptidase II (Lsp) inhibition is a viable strategy for studying lipoprotein processing.
  • This method provides insights into the complex posttranslational modification of bacterial lipoproteins.